Ohara J, Lahet S, Inman J, Paul W E
J Immunol. 1985 Oct;135(4):2518-23.
BSF-1 was partially purified from serum-free culture supernatants of cells of the EL-4 thymoma line, which had been induced 48 hr earlier with 4 beta-phorbol-12 beta-myristate-12 alpha-acetate (PMA). BSF-1 in 10-liter batches was adsorbed onto and eluted from trimethylsilyl-controlled pore glass beads (TMS-CpG) and then subjected to reverse-phase high-performance liquid chromatography (RP-HPLC). The recovery of BSF-1 activity by TMS-CpG and RP-HPLC ranged from 52 to 55% and 187 to 227%, respectively. The specific activity in units per milligram of protein of partially purified BSF-1 was approximately 2600 times higher than that of the culture supernatant protein. The partially purified BSF-1 had a single isoelectric point of 6.3 and an apparent m.w. between 18,000 and 21,700 when analyzed by isoelectric focusing and gel filtration-HPLC, respectively. The ability to prepare large amounts of partially purified BSF-1 by a rapid and efficient procedure should be of great help in both biochemical and immunologic studies of this lymphokine.
BSF-1是从EL-4胸腺瘤细胞系的无血清培养上清液中部分纯化得到的，该细胞系在48小时前用4β-佛波醇-12β-肉豆蔻酸酯-12α-乙酸酯（PMA）诱导。将10升批次的BSF-1吸附到三甲基硅烷基控制孔径玻璃珠（TMS-CpG）上并进行洗脱，然后进行反相高效液相色谱（RP-HPLC）。通过TMS-CpG和RP-HPLC回收的BSF-1活性分别为52%至55%和187%至227%。部分纯化的BSF-1每毫克蛋白质的比活性比培养上清液蛋白质高约2600倍。当分别通过等电聚焦和凝胶过滤-HPLC分析时，部分纯化的BSF-1具有单一的等电点6.3，表观分子量在18,000至21,700之间。通过快速有效的方法大量制备部分纯化的BSF-1的能力对于该淋巴因子的生化和免疫学研究应该有很大帮助。
